LOCUS       sequence-384890-        6267 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible Expression of N-Terminal 3X Flag Mouse C325A Caspase-9 
            Minus Large Subunit Region Val 177 to Arg 263.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6267)
  AUTHORS   Han J, Hou W, Goldstein LA, Stolz DB, Watkins SC, Rabinowich H
  TITLE     A Complex between Atg7 and Caspase-9: A NOVEL MECHANISM OF 
            CROSS-REGULATION BETWEEN AUTOPHAGY AND APOPTOSIS.
  JOURNAL   J Biol Chem. 2014 Mar 7;289(10):6485-6497. doi: 
            10.1074/jbc.M113.536854. Epub 2013 Dec 20.
  PUBMED    24362031
REFERENCE   2  (bases 1 to 6267)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1074/jbc.M113.536854"; journalName: "J Biol Chem"; date: 
            "2014-03-7- 7"; volume: "289"; issue: "10"; pages: "6485-6497"
COMMENT     Sequence Label: Tet-inducible 3xFlag Mouse C325A Caspase-9 Delta-A 
            LS
FEATURES             Location/Qualifiers
     source          1..6267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             996..1061
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2275..2292)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2281..2505
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2551..2979
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2638..2657)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2848..2869
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2988..3008)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2993..3322
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3173..3308
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3235..3254
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3370..3417
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3436..3810
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    3940..4061
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3977..3996)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4031..4050
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4110..4126)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4110..4126)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4123..4145)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4134..4150
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4158..4164,4165..4182,4183..4188))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4203..4224
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4341..4358)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4512..5100)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4592..4611)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5271..6062,6063..6131))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5894..5913
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6132..6236)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtaccg ccatggacta caaagaccat gacggtgatt
     1021 ataaagatca tgacatcgat tacaaggatg acgatgacaa gatggacgag gcggaccggc
     1081 agctcctgcg gcgatgcagg gtgcgcctag tgagcgagct gcaagtcgcg gagctctggg
     1141 acgctctgct gagtcgagag ctcttcacgc gcgacatgat cgaggatatt cagcaggcag
     1201 gatctgggtc tcggcgggat caggccaggc agctggtcac agaccttgag acccgaggga
     1261 ggcaggccct tcctctcttc atctcctgct tagaggacac aggccaaggc accctggctt
     1321 cactcttgca aagcggtcgg caagcagcca agcaggatcc agaggctgtt aaacccctag
     1381 accacctggt gcctgtggtc ctgggaccaa tgggactcac agcaaaggag cagagagtag
     1441 tgaagctgga cccgtcacag cctgccgtgg gaaacctcac cccagtggtg ctggggccag
     1501 aagagctctg gcctgctcgg ctcaagccag aggttctcag accagaaaca cccaggcccg
     1561 tggacattgg ttctggcgga gctcatgatg ccctggactg ctttgtggtg gtcatcctct
     1621 ctcatggctg ccaggccagc cacctccagt tcccgggtgc tgtctatggg acagatggat
     1681 gctccgtgtc cattgagaaa attgtgaata tcttcaacgg gagcggctgc cccagcctgg
     1741 gagggaagcc caagctcttc ttcatccagg ccgcgggtgg tgagcagaaa gaccatggct
     1801 ttgaggtggc ctgcacttcc tctcaaggca ggaccttgga cagtgactct gagccagatg
     1861 ctgtccccta tcaggaaggc ccaaggccct tggaccagct ggatgctgtg tcaagtttgc
     1921 ctacccccag tgacatcctt gtgtcctact ccaccttccc aggttttgtc tcctggaggg
     1981 acaagaaaag tggctcctgg tacatcgaga ccttggatgg cattctggag cagtgggctc
     2041 gctctgaaga cctgcagtcc ctccttctca gggttgccaa tgctgtttct gcgaaaggga
     2101 cttacaagca gattcctggc tgttttaact tcctccggaa aaagctgttt tttaaaactt
     2161 catgaggctc ggggctgctc tctgcctcat tattctctcc gaaggaattc tgcagatatc
     2221 cagcacagtg gcggccgctc gagtctagag ggcccgttta aacccgctga tcagcctcga
     2281 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     2341 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     2401 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     2461 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     2521 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     2581 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     2641 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     2701 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     2761 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     2821 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     2881 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     2941 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     3001 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3061 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3121 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3181 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3241 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     3301 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     3361 tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag
     3421 gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac
     3481 gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag
     3541 gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac
     3601 caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac
     3661 gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc
     3721 gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc
     3781 gtgcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc
     3841 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     3901 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     3961 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     4021 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     4081 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4141 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4201 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4261 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4321 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4381 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4441 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4501 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4561 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4621 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4681 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4741 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4801 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4861 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4921 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4981 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5041 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5101 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5161 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5221 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5281 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5341 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     5401 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     5461 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5521 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5581 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5641 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5701 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     5761 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     5821 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     5881 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     5941 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6001 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6061 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6121 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6181 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6241 tttccccgaa aagtgccacc tgacgtc
//
